Increasing Absorption Range for Photosynthesis http://media-2.web.britannica.com/eb-media/30/27030-004-293E0372.jpg
Goals Express different photosynthetic pigments in cyanobacteria Increase the wavelengths that can absorb light energy and transfer to photosynthetic machinery Potentially increase efficiency? http://www.emc.maricopa.edu/faculty/farabee/BIOBK/pigment.gif
Photosynthetic Pigments Chlorophylls Phycobilins Β carotene Carotenoids http://www.food-info.net/uk/colour/chlorophyll.htm http://www.mbari.org/staff/conn/botany/reds/daisy/phycocyanobilin.gif http://upload.wikimedia.org/wikipedia/commons/d/da/Beta-carotene.png
How do the Photo-Pigs work? Chlorophylls Transfers electrons around → reaction center Incorporated into photosystem protein complexes Carotenoids Membrane bound Transfers electrons to chlorophyll Phycobilins Water soluble, in cytoplasm Also transfers electrons to chlorophyll
Model Organism: Cyanobacteria Synechococcus, Synechocystis sp. PCC6803 Chlorophyll a, Phycobilins (phycocyanin, allophycocyanin, allophycocyanin B, phytoerythrin) Genetically tractable
What has been done before? Expressed chlorophyll b in Synechocystis sp. PCC6803 Demonstrated that it is incorporated into P700 chlorophyll-protein complex Satoh et al., Journal of Biological Chemistry, 2001 276 (6)
What can we do?
Idea 1: BioBrick: Bba_K152005 Four enzymes required for beta-carotene synthesis: geranylgeranyl diphosphate synthase (CrtE - BbaK152001) phytoene synthase (CrtB - BbaK152002) phytoene dehydrogenase (CrtI - BbaK18003) lycopene beta-cyclase (CrtY - BbaK152003)
Carotenoid Biosynthesis in Synechococcus
Questions to ask / Things to do When overexpressed is beta-carotene incorporated into a photosystem? What does beta-carotene normally do in cyanobacteria? How/Why was this biobrick made? What did they feed the E. coli? Can E. coli constructs be used in cyanobacteria?
Idea 2: Express Chlorophyll d Spectrum is red shifted Can act as reaction center? Need to find protein that can do this reaction. Larkum and Kuhl, Trends in Plant Science (2005) 10 (8).